Estimation of Shatavarin IV and Sarsasapogenin from the roots of Asparagus racemosus Wild using validated HPLC-ELSD method optimized using QbD approach

Abstract

The plant Asparagus racemosus Wild is mentioned in Ayurveda and used commercially alone or as an ingredient of many poly-herbal formulations indicated as a tonic and to improve the functioning of the reproductive system. Shatavarin type of saponin glycosides present in the roots were found to be responsible for the activity. An analytical method was developed using HPLC equipped with an evaporating light scattering detector (ELSD) to estimate Shatavarin-IV and a non-sugar moiety of this glycoside Sarsasapogenin from the dried roots of Asparagus racemosus. Chromatographic separation was achieved using Phenomenex^TM (C₁₈, 250 × 4.6 mm, id: 5 μm) column using an isocratic mobile phase comprising of methanol: water (95:05, %v/v) at a flow rate of 0.8 mL/min. The failure modes were identified and studied for their impact on Critical Method Attributes, and further, the chromatographic parameters were optimized using the design of the experiment approach. The developed method was found to be linear in the concentration range of 90–300 ng/mL for Shatavarin-IV and 150–500 ng/mL for Sarsasapogenin. The validation studies confirmed the precision, accuracy, and robustness of the developed analytical method. The plant material was found to contain 0.23 ± 0.01% w/w Shatavarin-IV and 1.48 ± 0.03% w/w Sarsasapogenin on a dried weight basis when estimated using the developed method.

Graphical Abstract

Keywords:

• Analytical DoE
• Asparagus racemosus
• HPLC-ESLD
• Sarsasapogenin
• Shatavarin-IV

Author contributions

Nishit Patel: Writing Original Draft, Investigation, Validation, Methodology, Data curation.

Manish Nandpal: Investigation, Validation, Review & Editing.

Archita Patel: Methodology, Review & Editing.

Samir Patel: Visualization, Review & Editing.

Manan Raval: Conceptualization, Project administration, Investigation, Supervision, Funding acquisition, Review & Editing.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This work was financially supported by the CHARUSAT seed research project sanctioned [CHARUSAT SEED RESEARCH GRANT/RPCP/MAR] to Dr. Manan Raval.